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==Procedure== | ==Procedure== | ||
# Combine sense and antisense | # Combine 5 μL sense oligonucleotide and 5 μL antisense oligonucleotide. Vortex and spin. | ||
# Double seal tube airtight with | # Double seal tube airtight with parafilm and press firmly in weighted holder. | ||
# Boil 800 mL water in a 1L beaker. | # Boil 800 mL water in a 1L beaker. | ||
# | # Wait 1 min. | ||
# Place weighted holder with sealed oligonucleotides into the beaker. | # Place weighted holder with sealed oligonucleotides into the beaker. | ||
# Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration. | # Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration. |
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